摘要
目的:为了利用流产儿角膜组织体外培养获取角膜上皮细胞,做为构建组织工程化角膜上皮的种子细胞。方法:收集30例(60个角膜)人流产胎儿,分别采用酶消化法、酶消化法结合组织块法和组织块法3种方法分离并培养人胎儿角膜上皮细胞。结果:用Dispase和胰蛋白酶冷消化角膜上皮后,在获取细胞总数、细胞活力和原代培养成功率上没有明显差异,但用Dispase消化后原代细胞容易成活,传代可得到纯化的上皮细胞;酶消化法结合组织块法培养时加入角膜缘组织块可以缩短细胞形成单层的时间,增加传代后细胞的活力;组织块法培养时,在胎儿全角膜组织培养时可得到角膜上皮细胞,角膜组织切割后培养不能获得纯化的角膜上皮细胞。结论:体外构建组织工程化角膜上皮时,种子细胞的获取可以采用流产儿角膜组织为材料分离并培养角膜上皮细胞。
AIM： To accumulate experiment data for isolation and culture of human corneal epithelial cells from abortive fetus and construction of tissue engineered corneas.
METHODS. Human corneal epithelial cells was isolated and cultivated from 30 abortive fetus （60 corneas） in vitro by enzyme digestion, tissue block method and tissue- block digested with enzyme method.
RESULTS： The results showed total cells, cell vitality and success rates of primary culture were not significantly different between Dispase and tripsin digestion, whereas the survival of primary cells digested with Dispase were easier contrast to tripsin and could obtain pure corneal epithelial cells. Monolayer forming time was shorten and vitality of passage cell was increased in tissue-block digested with enzyme method. Human pure corneal epithelial cells could be obtained when whole-corneas were cultured, but not when corneas tissue incised.
CONCLUSION： Human abortive fetus cornea can use as experimental material to isolate and culture human corneal epithelial cells.
出处
《国际眼科杂志》
CAS
2009年第2期250-253,共4页
International Eye Science
基金
中国国家自然科学基金项目(No.39970363)
中国国家重点基础研究发展规划项目(No.G1999054300)~~
关键词
人胎儿
角膜细胞
分离
培养
abortive fetus
corneal cells
isolate
cultivate
